**Human IFN-α ELISA Kit – For the Quantitative In Vitro Determination of Human Interferon α Concentrations in Serum, Plasma, Cerebrospinal Fluid, Tissue Homogenate, and Other Body Fluids**
**For Laboratory Research Use Only. Not for Diagnostic or Therapeutic Procedures.**
This kit is designed for the quantitative measurement of Human Interferon α (IFN-α) in various biological samples. The assay is based on a sandwich ELISA format, where the color change resulting from the reaction is measured at 450 nm using a microplate reader. A standard curve is generated using a set of calibration standards, allowing for accurate quantification of IFN-α levels in test samples.
**Key Features:**
- **Sensitivity:** <1.0 pg/mL
- **Dynamic Range:** 1.25–40 pg/mL
- **Intra- and Inter-assay CV <15%**
- **Cross-reactivity:** No significant cross-reactivity with other interferons or related proteins
**Sample Collection and Storage:**
- **Serum:** Use serum separator tubes. Allow clotting for 2 hours at room temperature or overnight at 4°C. Centrifuge at 2000×g for 20 minutes. Store at -20°C.
- **Plasma:** Collect using heparin as an anticoagulant. Centrifuge within 30 minutes at 2000×g for 30 minutes. Store at -20°C.
- **Tissue Homogenate, Cell Culture Supernatants, and Other Fluids:** Centrifuge to remove particulates. Assay immediately or store at -20°C. Avoid hemolysis and repeated freeze-thaw cycles.
**Materials Required (Not Supplied):**
- 37°C incubator
- Microplate reader (450 nm)
- Precision pipettes, disposable tips, and absorbent paper
- Distilled or deionized water
**Reagents Provided (Stored at 2–8°C):**
- Microtiter Strip Plate (12×8 or 12×4 strips)
- Standards (6 vials, 0.5 mL/vial)
- Sample Diluent (6.0 mL or 3.0 mL)
- HRP-Conjugate Reagent (10.0 mL or 5.0 mL)
- 20X Wash Solution (25 mL or 15 mL)
- Chromogen Solutions A & B (6.0 mL or 3.0 mL each)
- Stop Solution (6.0 mL or 3.0 mL)
- Closure Plate Membrane (2 units)
- User Manual and Sealed Bags
**Preparation Instructions:**
- Bring all reagents to room temperature (20–25°C) before use.
- Prepare 1X Wash Solution by diluting 1 volume of 20X Wash Solution with 19 volumes of distilled/deionized water.
- Do not use water baths for thawing; avoid using reagents beyond their expiration date.
**Assay Procedure:**
1. Prepare all reagents and add 50 µL of standard or sample to appropriate wells (excluding blank).
2. Cover with adhesive strip and incubate for 60 minutes at 37°C.
3. Wash the plate 4 times manually or automatically with 1X Wash Solution.
4. Add 50 µL of Chromogen A and B to each well. Incubate for 15 minutes at 37°C, protecting from light.
5. Add 50 µL of Stop Solution to each well. Read OD at 450 nm.
6. Plot the standard curve using average OD values and calculate sample concentrations accordingly.
**Important Notes:**
- Always perform a standard curve for each run.
- Ensure proper pipetting technique and consistent incubation conditions.
- Handle all biological materials as potentially infectious.
- Dispose of waste following local biosafety regulations.
- Store unused strips in sealed bags with desiccant at 2–8°C.
**Storage Conditions:**
- Short-term: 2–8°C (for frequent use)
- Long-term: -20°C (up to 6 months)
**Limitations:**
- This kit is not intended for diagnostic use.
- Results may vary due to operator technique, incubation time, or reagent age.
**Note:** If sample values exceed the highest standard, dilute with Sample Diluent and repeat the assay. Always follow good laboratory practices when handling biological samples.